LOCUS       sequence-63478-e        7140 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7140)
  AUTHORS   Zordan RE, Ren Y, Pan SJ, Rotondo G, De Las Penas A, Iluore J, 
            Cormack BP
  TITLE     Expression plasmids for use in Candida glabrata.
  JOURNAL   G3 (Bethesda). 2013 Oct 3;3(10):1675-86. doi: 10.1534/g3.113.006908.
  PUBMED    23934995
REFERENCE   2  (bases 1 to 7140)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: "10.1534/g3.113"; 
            journalName: "G3 (Bethesda)"; date: "2013-10-3- 3"; volume: "3"; 
            issue: "10"; pages: "1675-86"
COMMENT     Sequence Label: pCU-LYS21-GFP
FEATURES             Location/Qualifiers
     source          1..7140
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(10..28)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        666..737
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(738..806,807..1598)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(956..975)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1769..2357
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2258..2277
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2511..2528
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2645..2666
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2681..2686,2687..2704,2705..2711)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2719..2735
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     2724..2746
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2743..2759
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2743..2759
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2778..2798
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        2780..2798
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     CDS             4010..4726
                     /codon_start=1
                     /product="yeast-enhanced green fluorescent protein"
                     /label=yeGFP
                     /note="codons optimized for translation in S. cerevisiae 
                     and C. albicans"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(5156..5175)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5157..5175)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5185..5202)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5185..5201)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5194..5216)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5342..5797
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5429..5448)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5639..5660
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(5928..6731)
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     promoter        complement(6732..6947)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(6971..6990)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7090..7112
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
       61 cttagcaggt tagactaaca tgcaaaaagt ttatattcat ttttgtagtt attcttgttt
      121 tcattaggat gattacacac tgtggatgtg gattaatcga tcattgttaa tttagtgtag
      181 ctactaccat ttcaaacaaa acgtaaattc ttgcagtcgt actggatctg tgaatctatt
      241 agtatatatg aattaaagta gcttgtacat attattctgt tgaatcatat cgcagagcat
      301 tggttgaaat ccaaaatata aaaatgtaat atcacaaaaa ataatactaa ttctaacatt
      361 aatgggtcag atttttagtg aatacttaaa tttataatat cgtctattta agctagcaaa
      421 tggaacaaca tttaaagtaa gaacatcata tctacatgaa aatgtatatt tcaatctgac
      481 taataacgca gagcacatct ttcagtgatg tctgtcacat gatcaaaaag aattgtattt
      541 aaataattca taataaaagc ttaaaaaatt acaataatga aaataaagta aataatgaca
      601 tgggtaagag ttccgaaata gaatcttagt gtacaaaaca aaaattgcat cattagagat
      661 ccccacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
      721 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
      781 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
      841 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
      901 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
      961 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     1021 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     1081 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     1141 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     1201 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     1261 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     1321 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     1381 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     1441 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     1501 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     1561 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     1621 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     1681 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     1741 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     1801 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     1861 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     1921 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     1981 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     2041 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     2101 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     2161 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     2221 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     2281 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     2341 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     2401 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     2461 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     2521 tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga
     2581 ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg
     2641 caattaatgt gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg
     2701 ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc
     2761 atgattacgc caagcgcgca attaaccctc actaaaggga acaaaagctg gagctcagaa
     2821 agcgaagaag atatatctag tttttctccg cactcttacc cattctcatt ttttttaact
     2881 cagcatttta ctatagattt tttttctttc ttataatcaa gtacatagaa tatatgcaca
     2941 tagaaacaaa aaagataggc aagttattat tcagacgcat tatcataaaa aactttattt
     3001 atctgttaac gaaacaaaat ttagcgctat ttctcaaagg attgtagcag catcatgaga
     3061 aaaatatcgt caatcggagc gaagtttatt acgtctacac aggtaacaag ggtatctgtg
     3121 tattgaacct cggattgaac cggcatattc tcacacaaat gagaagcttg aaatccccac
     3181 ggatcctgcc acacccccaa caattaaagt gactaagaaa caactctaga cccttgttac
     3241 ttatttcact caaacttgta cgtgccctgc taacacagtt tacagatagg agtccatatg
     3301 actgcaattc cacacacctc gaatggctct tatcctctcc cctctccaat agaagagact
     3361 tgagctaaga aaaaatttac atcggaattt ttacagctgt gttggtttca tcactacttt
     3421 gcaagcctag tttttaccac ttcctcatat ggttatgact cttttcctca tctttttggt
     3481 gctttcattt ctttttgcaa cggtttataa tttttacctt cgcaattttc tgattttttt
     3541 ttgcaaaaat attttcacaa tttacaaaat gcactaaaag tccgtagaga agtgggatca
     3601 gttattattc cttcacttac taacatcgat acatttcgaa tcagctattt catagtggtt
     3661 tatgctattc taatcacaca ttttgttaca acacttcctt caatacacga ttgtagttgt
     3721 tgtacgtatc catatagtcg actcatttgc gtcgccttct cttgctttta gcaacagtcg
     3781 ttctagtttt tgcaagcttc tgactaattc ctaaaggcaa gtataaatac atgccgtttc
     3841 ttgctgaaac catatgttat ctcgtatcta tgtttcatta cacatttttg taacagctat
     3901 ttcaactggt gaaatactac aagtaaaaat atattctacc taaaatattt ttgctgaaca
     3961 aagaatatta aagactagaa ctagtggatc ccccgggctg caggaattca tgtctaaagg
     4021 tgaagaatta ttcactggtg ttgtcccaat tttggttgaa ttagatggtg atgttaatgg
     4081 tcacaaattt tctgtctccg gtgaaggtga aggtgatgct acttacggta aattgacctt
     4141 aaaatttatt tgtactactg gtaaattgcc ggttccatgg ccaaccttag tcactacttt
     4201 cggttatggt gttcaatgtt ttgctagata cccagatcat atgaaacaac atgacttttt
     4261 caagtctgcc atgccagaag gttatgttca agaaagaact atttttttca aagatgacgg
     4321 taactacaag accagagctg aagtcaagtt tgaaggtgat accttagtta atagaatcga
     4381 attaaaaggt attgatttta aagaagatgg taacatttta ggtcacaaat tggaatacaa
     4441 ctataactct cacaatgttt acatcatggc tgacaaacaa aagaatggta tcaaagttaa
     4501 cttcaaaatt agacacaaca ttgaagatgg ttctgttcaa ttagctgacc attatcaaca
     4561 aaatactcca attggtgatg gtccagtctt gttaccagac aaccattact tatccactca
     4621 atctgcctta tccaaagatc caaacgaaaa gagagaccac atggtcttgt tagaatttgt
     4681 tactgctgct ggtattaccc atggtatgga tgaattgtac aaataactgc aggtcgacct
     4741 cgagaacaca acccacagct accaccatca acaatattta tatatataac gtacacatag
     4801 aaatcacaca aacagagtat ttattcttaa ctacatgaac taccatcaga ccgtctgggc
     4861 cactatataa tgtgccattc ataaacgtga tcactttacg tagcaggcaa ccccaggtga
     4921 aaatttttca gcgagctgcc agattgtcag gtgaaaaact gaaaaaaact tctgggcgat
     4981 gagcttgtgg cgggaaatta agtatataaa gcagttagtt tcttctgctt cttgtggttc
     5041 tgggtattct tgtaatattt ggaagaatag aaaatcaaga ctactgcctt tcttttcata
     5101 ttatagagga agaaatacgc acgaacacga tatagaggta aaggtaccca attcgcccta
     5161 tagtgagtcg tattacgcgc gctcactggc cgtcgtttta caacgtcgtg actgggaaaa
     5221 ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa
     5281 tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg
     5341 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     5401 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     5461 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     5521 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     5581 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     5641 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     5701 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     5761 aacgcgaatt ttaacaaaat attaacgctt acaatttcct gatgcggtat tttctcctta
     5821 cgcatctgtg cggtatttca caccgcatag ggtaataact gatataatta aattgaagct
     5881 ctaatttgtg agtttagtat acatgcattt acttataata cagtttttta gttttgctgg
     5941 ccgcatcttc tcaaatatgc ttcccagcct gcttttctgt aacgttcacc ctctacctta
     6001 gcatcccttc cctttgcaaa tagtcctctt ccaacaataa taatgtcaga tcctgtagag
     6061 accacatcat ccacggttct atactgttga cccaatgcgt ctcccttgtc atctaaaccc
     6121 acaccgggtg tcataatcaa ccaatcgtaa ccttcatctc ttccacccat gtctctttga
     6181 gcaataaagc cgataacaaa atctttgtcg ctcttcgcaa tgtcaacagt acccttagta
     6241 tattctccag tagataggga gcccttgcat gacaattctg ctaacatcaa aaggcctcta
     6301 ggttcctttg ttacttcttc tgccgcctgc ttcaaaccgc taacaatacc tgggcccacc
     6361 acaccgtgtg cattcgtaat gtctgcccat tctgctattc tgtatacacc cgcagagtac
     6421 tgcaatttga ctgtattacc aatgtcagca aattttctgt cttcgaagag taaaaaattg
     6481 tacttggcgg ataatgcctt tagcggctta actgtgccct ccatggaaaa atcagtcaag
     6541 atatccacat gtgtttttag taaacaaatt ttgggaccta atgcttcaac taactccagt
     6601 aattccttgg tggtacgaac atccaatgaa gcacacaagt ttgtttgctt ttcgtgcatg
     6661 atattaaata gcttggcagc aacaggacta ggatgagtag cagcacgttc cttatatgta
     6721 gctttcgaca tgatttatct tcgtttcctg caggtttttg ttctgtgcag ttgggttaag
     6781 aatactgggc aatttcatgt ttcttcaaca ctacatatgc gtatatatac caatctaagt
     6841 ctgtgctcct tccttcgttc ttccttctgt tcggagatta ccgaatcaaa aaaatttcaa
     6901 ggaaaccgaa atcaaaaaaa agaataaaaa aaaaatgatg aattgaaaag gtggtatggt
     6961 gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa
     7021 cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg
     7081 tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga
//
